Synthesis and characterization of antimicrobial colloidal polyanilines.

The potential application of colloidal polyaniline (PANI) as an antimicrobial is limited by challenges related to solubility in common organic solvents, scalability, and antimicrobial potency. To address these limitations, we introduced a functionalized PANI (fPANI) with carboxyl groups through the polymerisation of aniline and 3-aminobenzoic acid in a 1:1 molar ratio. fPANI is more soluble than PANI which was determined using a qualitative study. We further enhanced the solubility and antimicrobial activity of fPANI by incorporating Ag nanoparticles onto the synthesized fPANI colloid via direct addition of 10 mM AgNO3. The improved solubility can be attributed to an approximately 3-fold reduction in size of particles. Mean particle sizes are measured at 1322 nm for fPANI colloid and 473 nm for fPANI-Ag colloid, showing a high dispersion and deagglomeration effect from Ag nanoparticles. Antimicrobial tests demonstrated that fPANI-Ag colloids exhibited superior potency against Gram-positive Staphylococcus aureus, Gram-negative Escherichia coli, and Bacteriophage PhiX 174 when compared to fPANI alone. The minimum bactericidal concentration (MBC) and minimum virucidal concentration (MVC) values were halved for fPANI-Ag compared to fPANI colloid and attributed to the combination of Ag nanoparticles with the fPANI polymer. The antimicrobial fPANI-Ag colloid presented in this study shows promising results, and further exploration into scale-up can be pursued for potential biomedical applications.

Optimizing the extraction process of natural antioxidants from chardonnay grape marc using microwave-assisted extraction.

The aim of this work was to extract phenolic compounds from Chardonnay grape marc employing a microwave-assisted extraction (MAE). Firstly, the effect of solvent concentration (30-60%), solid mass (1.0-2.0 g) and extraction time (5-15 min) on the recovery of phenolic content and antioxidant capacity was evaluated using a response surface methodology (RSM). The optimal parameters found by RSM were 48% ethanol for the solvent content, 10 min for the extraction time, and 1.77 g for the solid mass. The extraction was carried out at room temperature to increase scaling-up opportunities at industrial level. It was found that the phenolic profile was mainly composed of flavanols, such as procyanidins, catechin and epicatechin. Furthermore, the polyphenols obtained by MAE showed a DPPH· inhibition value of 87 ±â€¯5% and the total phenolic content was 1.21 ±â€¯0.04 mg GAE/mL. Finally, it was observed that the degradation temperature of the extract (≈ 200 °C) was above the temperature commonly used for the manufacture of protein films by thermo-mechanical processes. This highlights the potential use of this extract as a bioactive additive in protein film forming formulations for food and pharmaceutical applications.

The antimicrobial action of polyaniline involves production of oxidative stress while functionalisation of polyaniline introduces additional mechanisms.

Polyaniline (PANI) and functionalised polyanilines (fPANI) are novel antimicrobial agents whose mechanism of action was investigated. Escherichia coli single gene deletion mutants revealed that the antimicrobial mechanism of PANI likely involves production of hydrogen peroxide while homopolymer poly(3-aminobenzoic acid), P3ABA, used as an example of a fPANI, disrupts metabolic and respiratory machinery, by targeting ATP synthase and causes acid stress. PANI was more active against E. coli in aerobic, compared to anaerobic, conditions, while this was apparent for P3ABA only in rich media. Greater activity in aerobic conditions suggests involvement of reactive oxygen species. P3ABA treatment causes an increase in intracellular free iron, which is linked to perturbation of metabolic enzymes and could promote reactive oxygen species production. Addition of exogenous catalase protected E. coli from PANI antimicrobial action; however, this was not apparent for P3ABA treated cells. The results presented suggest that PANI induces production of hydrogen peroxide, which can promote formation of hydroxyl radicals causing biomolecule damage and potentially cell death. P3ABA is thought to act as an uncoupler by targeting ATP synthase resulting in a futile cycle, which precipitates dysregulation of iron homeostasis, oxidative stress, acid stress, and potentially the fatal loss of proton motive force.

Investigation of Polyaniline and a Functionalised Derivative as Antimicrobial Additives to Create Contamination Resistant Surfaces.

Antimicrobial surfaces can be applied to break transmission pathways in hospitals. Polyaniline (PANI) and poly(3-aminobenzoic acid) (P3ABA) are novel antimicrobial agents with potential as non-leaching additives to provide contamination resistant surfaces. The activity of PANI and P3ABA were investigated in suspension and as part of absorbent and non-absorbent surfaces. The effect of inoculum size and the presence of organic matter on surface activity was determined. PANI and P3ABA both demonstrated bactericidal activity against Escherichia coli and Staphylococcus aureus in suspension and as part of an absorbent surface. Only P3ABA showed antimicrobial activity in non-absorbent films. The results that are presented in this work support the use of P3ABA to create contamination resistant surfaces.

The tuberculocidal activity of polyaniline and functionalised polyanilines.

Tuberculosis is considered a leading cause of death worldwide. More than 95% of cases and deaths occur in low- and middle-income countries. In resource-limited countries, hospitals often lack adequate facilities to manage and isolate patients with infectious tuberculosis (TB), relying instead on personal protective equipment, such as facemasks, to reduce nosocomial transmission of the disease. Facemasks impregnated with an antimicrobial agent may be a cost-effective way of adding an extra level of protection against the spread of TB by reducing the risk of disease transmission. Conducting polymers, such as polyaniline (PANI), and their functionalised derivatives are a novel class of antimicrobial agents with potential as non-leaching additives to provide contamination resistant surfaces. We have investigated the antimicrobial action of PANI and a functionalised derivative, poly-3-aminobenzoic acid (P3ABA), against mycobacteria and have determined the optimal treatment time and concentration to achieve significant knockdown of Mycobacterium smegmatis and Mycobacterium tuberculosis on an agar surface. Results indicated that P3ABA is a potential candidate for use as an anti-tuberculoid agent in facemasks to reduce TB transmission.

Functionalized polyanilines disrupt Pseudomonas aeruginosa and Staphylococcus aureus biofilms.

The purpose of the present study was to investigate the antimicrobial effects of functionalized polyanilines (fPANIs) against stationary phase cells and biofilms of Pseudomonas aeruginosa and Staphylococcus aureus using homopolymer of sulfanilic acid (poly-SO3H) as a model. The chemically synthesized poly-SO3H was characterized using Fourier Transform Infra-Red (FTIR) and Ultraviolet-Visible (UV-Vis) spectroscopies. The molecular weight (Mw) and elemental analysis of homopolymer poly-SO3H were also examined. We found that poly-SO3H was bactericidal against stationary phase cells of P. aeruginosa and S. aureus at a concentration of 20 mgml(-1). Surprisingly, we discovered that the same concentration (20 mgml(-1)) of poly-SO3H significantly disrupted and killed bacterial cells present in pre-established forty-eight hour static biofilms of these organisms, as shown by crystal violet and bacterial live/dead fluorescence staining assays. In support of these data, poly-SO3H extensively diminished the expression of bacterial genes related to biofilm formation in stationary phase cells of P. aeruginosa, and seemed to greatly reduce the amount of the quorum sensing molecule N-(3-oxododecanoyl)-l-homoserine lactone (3OC12-HSL) able to be recovered from biofilms of this organism. Furthermore, we found that poly-SO3H was able to effectively penetrate and kill cells in biofilms formed by the P. aeruginosa (AESIII) isolate derived from the sputum of a cystic fibrosis patient. Taken together, the results of the present study emphasise the broad antimicrobial activities of fPANI, and suggest that they could be developed further and used in some novel ways to construct medical devices and/or industrial equipment that are refractory to colonization by biofilm-forming bacteria.

Influence of heat curing on structure and physicochemical properties of phenolic acid loaded proteinaceous electrospun fibers.

Effects of heat treatment on structure and physicochemical properties of zein (Ze) and gallic acid loaded zein (Ze-GA) electrospun fiber mats were investigated. The electrospun fiber mats displayed different surface and physicochemical properties after being heat-cured at 150 °C for 24 h, which were closely related to the initial amount of loaded gallic acid. The gallic acid was released from the Ze-GA fiber mats in a constant manner, but heat curing decreased the rate of release. Heat curing remarkably increased the molecular weight of the Ze and Ze-GA electrospun fiber mats. Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) analysis of the fiber mats indicated variations in zein protein secondary structure after heat curing. (13)C solid state NMR (SS-NMR) confirmed the presence of a different chemical environment among the fiber mats. The fabrication of heat-cured zein based electrospun fibers in this study may find applications in the food packaging industry.

Evaluation of gallic acid loaded zein sub-micron electrospun fibre mats as novel active packaging materials.

The applicability of gallic acid loaded zein (Ze-GA) electrospun fibre mats towards potential active food packaging material was evaluated. The surface chemistry of the electrospun fibre mats was determined using X-ray photon spectroscopy (XPS). The electrospun fibre mats showed low water activity and whitish colour. Thermogravimetric analysis (TGA) and Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy revealed the stability of the fibre mats over time. The Ze-GA fibre mats displayed similar rapid release profiles, with Ze-GA 20% exhibiting the fastest release rate in water as compared to the others. Gallic acid diffuses from the electrospun fibres in a Fickian diffusion manner and the data obtained exhibited a better fit to Higuchi model. L929 fibroblast cells were cultured on the electrospun fibres to demonstrate the absence of cytotoxicity. Overall, the Ze-GA fibre mats demonstrated antibacterial activity and properties consistent with those considered desirable for active packaging material in the food industry.

Encapsulation of food grade antioxidant in natural biopolymer by electrospinning technique: a physicochemical study based on zein-gallic acid system.

Gallic acid was successfully incorporated into zein ultra-fine fibres at different loading amount (5%, 10% and 20%) in order to develop an encapsulating technology for functional ingredient delivery using electrospinning. The produced fibres exhibit diameters ranging from 327 to 387 nm. The physical and thermal properties of encapsulated gallic acid were determined by X-ray diffraction (XRD) and differential scanning calorimetry (DSC); and the interaction between gallic acid and zein was attested by attenuated total reflection-Fourier transform infrared (ATR-FTIR). Thermogravimetric analysis (TGA) demonstrated a different thermal stability of the fabricated complex before and after the gallic acid incorporation. Lastly, the 1,1'-diphenyl-2-picrylhydrazyl (DPPH) assay showed that the gallic acid had retained its antioxidant activity after incorporation in zein electrospun fibres. Overall, electrospinning technique had shown promising results as an efficient and effective method for the preparation of sub-micron structured encapsulated functional ingredient that may find uses in food industry.

Broad spectrum antimicrobial activity of functionalized polyanilines.

The antimicrobial properties of conductive functionalized polyanilines (fPANI) were investigated by exploring their interaction with bacterial cells. In sharp contrast to polyaniline (PANI), lower concentrations of fPANI were needed to strongly inhibit the growth of wild-type Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus, as well as several antibiotic-resistant clinical pathogens. To gain an insight into how fPANI have an impact on cellular physiology we used a whole genome expression study in the model E. coli MG1655 strain exposed to a representative fPANI. The expression levels of 218 (∼5.1%) genes changed significantly. Moreover, we found that certain oxidative damage-responsive genes were strongly induced, while genes potentially involved in energy metabolism and transport and in forming bacterial cell walls and stress-resistant cellular communities (biofilms) were repressed. Taken together, our results appear to indicate that the antimicrobial effects of fPANI, in part at least, might stem from their ability to target the operations of multiple and diverse cellular processes, and suggest that fPANI could be useful ingredients for biomaterials used in the development of food packaging and medical devices.

Electrospun functionalized polyaniline copolymer-based nanofibers with potential application in tissue engineering.

Nanofibrous blends of HCl-doped poly(aniline-co-3-aminobenzoic acid) (3ABAPANI) copolymer and poly(lactic acid) (PLA) were fabricated by electrospinning solutions of the polymers, in varying relative proportions, in dimethyl sulfoxide/tetrahydrofuran mixture. The morphology, mechanical and electrical properties of the nanofibers were characterized and an assessment of their bioactivity performed. To assess cell morphology and biocompatibility, pure PLA and 3ABAPANI-PLA nanofibrous mats were deposited in the form of three-dimensional networks with a high degree of connectivity, on glass substrates, and their ability to promote proliferation of COS-1 fibroblast cells was determined. The nanofibrous electrospun 3ABAPANI-PLA blends gave enhanced cell growth, potent antimicrobial capability against Staphylococcus aureus and electrical conductivity. This new class of nanofibrous blends can potentially be employed as tissue engineering scaffolds, and in particular have showed promise as the basis of a new generation of functional wound dressings that may eliminate deficiencies of currently available antimicrobial dressings.

A rapid and facile synthesis of nanofibrillar polyaniline using microwave radiation.

We present the first fast and facile microwave assisted synthesis of polyaniline (PANI) nanofibers ("MWA synthesis"). Under conventional synthesis (CS), the polymer was produced with 79.7% yield after 5 h at ambient temperature. However, under microwave irradiation, the nanofibers were produced with yield of 76.2% after only 5 min, i.e., with 78.8% after 20 min at ambient temperature. The FTIR and Raman spectra show the PANI structure in all samples either synthesized conventionally or in the microwave. SEM and TEM confirm the nanofibrillar morphology.